Figure 5
WEF suppresses PMA-induced p38 and JNK phosphorylation as well as NF-κB activation.
(A): After treatment with or without WEF (100 μg/ml) for 12 h, cells were stimulated with PMA (5 nM) for 15, 30, or 60 min. Total cell lysates were subjected to Western blotting to evaluate phosphorylation of p38, ERK and JNK. Phosphorylation and degradation of IκBα were also measured. After normalization to tubulin, the relative ratios of phosphorylated protein/total protein were calculated. (B): Cells pre-treated with WEF (25, 50, or 100 μg/ml) for 12 h were stimulated with PMA for 30 min and then subjected to Western blotting. Total cell lysates were measured for IκBα phosphorylation and degradation. Cytosolic and nuclear fractions were prepared to evaluate nuclear translocation of the NF-κB p65 subunit after PMA stimulation. Tubulin and TBP were used as loading control for cytosolic and nuclear compartment, respectively. Data are expressed as the mean ± SD of two independent experiments. The full size blots were shown in the Supplementary Figure S4 and band of interest is indicated with an arrow. #, p<0.01 vs. untreated control. *, p<0.01 vs. PMA stimulation.
